CN102380102A - Method for preparing environment-responsive mesoporous silicon nanoparticles - Google Patents
Method for preparing environment-responsive mesoporous silicon nanoparticles Download PDFInfo
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- CN102380102A CN102380102A CN 201110342007 CN201110342007A CN102380102A CN 102380102 A CN102380102 A CN 102380102A CN 201110342007 CN201110342007 CN 201110342007 CN 201110342007 A CN201110342007 A CN 201110342007A CN 102380102 A CN102380102 A CN 102380102A
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Abstract
The invention relates to a method for preparing environment-responsive mesoporous silicon nanoparticles. The mesoporous silicon nanoparticles with the particle size of 20 to 1,000nm, the pore size of 2 to 50nm, the specific surface area of 500 to 1,500m<2>/g and a controlled structure are prepared by a template extraction method and subjected to surface functionalization by a self-assembly technology. A medicine sustained-release system prepared by the method is high in biocompatibility and medicine loading rate, can be released intelligently and controllably and has good application prospects in fields of growth factor and gene release, enzyme immobilization, cancer treatment and the like in tissue engineering, the preparation method is simple, reaction conditions are mild, and experiment raw materials are low in cost.
Description
Technical field
The invention belongs to the preparation field of mesoporous silicon nano, particularly a kind of environmental responsibility mesoporous silicon nanometer particle process method.
Background technology
In process of tissue reparation, the controlled release of somatomedin is raised, is regulated cell function and promote that organization healing has important function seed cell.Most of somatomedin are short and changeableness of half-life in vivo, and excessive release maybe toxigenicity and carcinogenecity.Therefore, setting up effective controlled release carrier is the essential condition that realizes its biological function.In organizational project, adopt two kinds of different somatomedin mode of loadings usually: (1) chemical method is fixed to somatomedin on the carrier, combines through covalent bond, hydrogen bond or electrostatic interaction between somatomedin and the carrier; (2) method of physically trapping is coated on somatomedin in polymer microballoon or the gel, comes the slow release of the control growing factor through diffusion or degraded.Therefore, make up nanoscale and micron-sized drug release carrier and to combine with support be a Critical policies of realization somatomedin sustained release.
Current, cancer remains one of killer who threatens human health, and the treatment of cancer great difficult problem that also to be medical circle face.Main cause is: the certain toxic and side effects of (1) existing cancer therapy drug ubiquity, human normal tissue is brought injury; (2) there is very big defective in traditional administering mode self, and bioavailability of medicament is relatively poor; (3) multidrug resistance of tumor causes chemotherapy failure and disease relapse.Analysis according to statistics, 90% malignant tumor patient is died from the multidrug resistance of tumor.Therefore the reverse characteristic of multidrug resistance is present oncotherapy research focus.Common resolution policy comprises: combine the chemical sensitization medicine to rebuild the main medicine of imitating, the applying biological nanotechnology improves operational efficiency.Said method combined can provide efficacious therapy means.Therefore, the applying nano technology is constructed the drug-loading system of anticarcinogen, chemical sensitization medicine, can play an important role aspect increase drug effect, the reverse multidrug resistance.Usually, these nano medicament carrying systems are mainly based on the organic or inorganic nano-carrier, for example liposome, polymer micelle and silicon, carbon meso-porous nano material etc.Though existing drug delivery system has certain drug loading, good stable property and biocompatibility, existence is little like easy leakage in the drug delivery process, useful load, is difficult to obtain drawbacks such as controllable sustained-release.Therefore improve drug loading, the toxic and side effects of reduction cancer therapy drug realizes the medicine intelligent control, is the key that effectively realizes treatment of cancer.
Mesoporous silicon nano has application widely aspect the structure medicine intelligence slow-released carrier.Mesoporous silicon has good biocompatibility, specific surface area is big, specific pore volume is big, the aperture structure homogeneous can be in harmonious proportion characteristics such as the easy modification in surface, has special advantages as medicine and gene controlled release carrier.Be rich in the characteristics that the silicon hydroxyl is easy to modify according to mesoporous silicon outer surface, can design the mesoporous silicon medicine-carried system of various environment-responsives.People utilize various chemical entities to comprise that conducts " gate inhibition " such as nanoparticle, organic molecule, super-molecule assembling body carry out sealing of hole or add medicated cap mesoporous silicon; These chemical entities can stimulate like light, temperature, pH value, Reducing agent and enzyme etc. to external world makes quick response; And then the opening in control duct with close, realize being carried on the controlling slow release of the object drug molecule in the duct.Existing mesoporous silicon medicine-carried system respectively has pluses and minuses.Form hollow microcapsule like parcel polyelectrolyte on colloidal particle, can realize the drug release of pH response, but pore passage structure is prone to subside in the preparation process, be not suitable for producing in enormous quantities; Tubulose mesoporous material drug-loading system has bigger drug loading, but rate of release is difficult to control.Therefore, in conjunction with the advantage of mesoporous silicon, has application promise in clinical practice undoubtedly through prepared by surface modification environment-responsive drug delivery system.For example, amycin etc. are a kind of broad-spectrum antitumor drug, and it can produce biochemical effect widely to body, but stronger cytotoxicity is arranged.Adopting the environment-responsive slow-released carrier is the ideal selection of this type of medicine of controlled release.
Summary of the invention
Technical problem to be solved by this invention provides a kind of environmental responsibility mesoporous silicon nanometer particle process method, and the mesoporous silicon nano drug loading of pH response medicine carrying of this method preparation is high, can intelligent controlled release, have excellent biological compatibility.
A kind of environmental responsibility mesoporous silicon nanometer particle process method of the present invention comprises:
(1) template is dissolved in the sodium hydroxide solution under 50-95 ℃, adds ethyl orthosilicate, stir, sucking filtration is dry, alcohol reflux go the template final vacuum dry mesoporous silicon nano; Wherein, the mol ratio of template, sodium hydroxide and ethyl orthosilicate is 1-2: 3-5: 7-9;
(2) above-mentioned mesoporous silicon is scattered in the toluene, adds the 3-aminopropyltriethoxywerene werene, reaction under nitrogen protection, dry must amido modified mesoporous silicon nano; Wherein, the mol ratio of mesoporous silicon, 3-aminopropyltriethoxywerene werene and toluene is 4-6: 0.5-2: 400-600;
(3) the amido modified mesoporous silicon nano that mesoporous silicon nano that step (1) is made or step (2) make is processed suspension, adds anionic polyelectrolyte or cationic polyelectrolyte solution, stirs 0.2-2h, and centrifuge washing disperses; Repeat this process after drying, obtain the mesoporous silicon nano of the autonomous finishing decorations in surface;
(4) bioactive molecule is placed the buffer solution of pH 2.0-8.0, add the mesoporous silicon nano that decorations are independently fitted up on above-mentioned surface, stir 4-20h, again pH is transferred to 6.0-8.0, stir 0.2-2h, centrifuge washing is dry, promptly gets.
Template in the said step (1) is a cetyl trimethyl ammonium bromide.
The particle diameter of the mesoporous silicon in the said step (1) is 20-1000nm, and specific surface area is 500-1500m
2/ g, average pore size is 2-50nm.
Anionic polyelectrolyte in the said step (3) is maleic anhydride sodium salt, sodium alginate, kayexalate, polyacrylic acid, polymethylacrylic acid or polyacrylamide; Cationic polyelectrolyte is chitosan, polyallylamine hydrochlorate, PDDA or heparin.
Anionic polyelectrolyte or cationic polyelectrolyte solution concentration in the said step (3) are 0.2-2mg/mL.
Bioactive molecule in the said step (4) is somatomedin, gene, enzyme or cancer therapy drug.
The buffer solution of pH 2.0-8.0 is pH 2.0-3.0 in the said step (4), hydrochloric acid solution or pH 3.0-6.0, acetate buffer solution or pH 6.0-8.0, phosphate buffer.
Said somatomedin is phosphoric acid sphingol, VEGF, basic fibroblast growth factor or epidermal growth factor.
Said gene is microRNA, disturb little RNA, messenger RNA, transfer RNA or ribosomal RNA.
Said enzyme is pectase, protease, lipase, catalase or amylase.
Said anticarcinogen is amycin, dexamethasone, lomustine, carmustine, dexrazoxane, fludarabine phosphate, capecitabine, paclitaxel, replaces lucky Austria, oxaliplatin, docetaxel, vinorelbine, elemene or hydroxy camptothecin.
Beneficial effect
(1) the present invention is simple to operate, reaction condition is gentle, and experimental raw is cheap, good economy performance, good biocompatibility;
(2) the mesoporous silicon nano drug loading of pH response medicine carrying of the present invention's preparation is high; Can intelligent controlled release, have excellent biological compatibility, be applicable to micromolecule cancer therapy drug, gene and somatomedin targeting and sustained release; Stable performance also is easy to preserve, and has a good application prospect.
Description of drawings
Fig. 1 is the field emission scanning electron microscope photo of embodiment 1,2 products therefroms;
Fig. 2 is the transmission electron microscope photo of embodiment 1,2 products therefroms;
Fig. 3 is the zeta potential figure of embodiment 1,2 gained self assembling processes;
Fig. 4 is the growth factor slow-release curve of embodiment 1 products therefrom;
Fig. 5 is the anti-cancer medicine slow release curve of embodiment 2 products therefroms.
The specific embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in the restriction scope of the present invention.Should be understood that in addition those skilled in the art can do various changes or modification to the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Embodiment 1
The pH responsive nano particle drug delivery system of adjustable release phosphoric acid sphingol
(1) the template extraction method prepares mesoporous silicon nano, and the template cetyl trimethyl ammonium bromide is dissolved in the sodium hydroxide solution under 50 ℃, dropwise adds ethyl orthosilicate; Vigorous stirring produces white precipitate, and sucking filtration is dry, after alcohol reflux 8h removes template; Vacuum drying is preserved subsequent use;
(2) the mesoporous silicon with step 1 preparation is dispersed in the toluene; Add the 3-aminopropyltriethoxywerene werene; The mol ratio of silicon dioxide, 3-aminopropyltriethoxywerene werene and toluene is 5: 1: 500; Reaction 20h under nitrogen protection, dry amido modified mesoporous silicon nano is preserved subsequent use;
(3) the amido modified mesoporous silicon of step 2 preparation is processed suspension (1.0-2.0wt.%), add solution of sodium alginate (1-2mg/mL), stirring at room 0.2-2h, the centrifugal 3min of 4000r/min; Washing is dispersed into suspension, adds chitosan solution (1-2mg/mL), stirring at room 0.2-2h; The centrifugal 3min of 4000r/min, washing disperses to process suspension; Whole process repeats 4 times, obtains the mesoporous silicon nano of the autonomous finishing decorations in surface, drying for standby;
(4) with the mesoporous silicon nano of the finishing of step 3 preparation, add in the phosphate buffer solution (pH6.5-8.0) of phosphoric acid sheath saddle alcohol, stir 10-24h and carry out medicine absorption.Again pH is transferred to 5.5-6.5, stir 2h, centrifuge washing is dry.Obtain the responsibility mesoporous silicon nano of pH of loaded with growth.
Preparation loads the pH responsive nano particle of amycin
(1) the template extraction method prepares mesoporous silicon nano, and the template cetyl trimethyl ammonium bromide is dissolved in the sodium hydroxide solution under 95 ℃, dropwise adds ethyl orthosilicate; Vigorous stirring produces white precipitate, and sucking filtration is dry, after alcohol reflux 24h removes template; Vacuum drying is preserved subsequent use;
(2) the mesoporous silicon nano of step 1 preparation is processed suspension (1.0-2.0wt.%), add polyallylamine hydrochlorate (1-2mg/mL), stirring at room 0.2-2h, the centrifugal 3min of 4000r/min; Washing disperses to process suspension, adds kayexalate solution (1-2mg/mL), stirring at room 0.2-2h; The centrifugal 3min of 4000r/min, washing disperses to process suspension; Whole process repeats 4 times, independently adorns drying layer by layer;
(3) with the mesoporous silicon of the parcel of step 2 preparation; In the phosphate buffer solution of micromolecule cancer therapy drug pH 1.5-4.5 such as adding amycin, stir 10h, carry out medicine to load absorption; Again pH is transferred to 6.5-8.0; Stir 2h, centrifuge washing is dry, promptly gets the mesoporous silicon nano of the pH response of loading small-molecule drug.
Claims (10)
1. environmental responsibility mesoporous silicon nanometer particle process method comprises:
(1) template is dissolved in the sodium hydroxide solution under 50-95 ℃, adds ethyl orthosilicate, stir, sucking filtration is dry, alcohol reflux go the template final vacuum dry mesoporous silicon nano; Wherein, the mol ratio of template, sodium hydroxide and ethyl orthosilicate is 1-2: 3-5: 7-9;
(2) above-mentioned mesoporous silicon is scattered in the toluene, adds the 3-aminopropyltriethoxywerene werene, reaction under nitrogen protection, dry must amido modified mesoporous silicon nano; Wherein, the mol ratio of mesoporous silicon, 3-aminopropyltriethoxywerene werene and toluene is 4-6: 0.5-2: 400-600;
(3) the amido modified mesoporous silicon nano that mesoporous silicon nano that step (1) is made or step (2) make is processed suspension, adds anionic polyelectrolyte or cationic polyelectrolyte solution, stirs 0.2-2h, and centrifuge washing disperses; Repeat this process after drying, obtain the mesoporous silicon nano of the autonomous finishing decorations in surface;
(4) bioactive molecule is placed the buffer solution of pH 2.0-8.0, add the mesoporous silicon nano that decorations are independently fitted up on above-mentioned surface, stir 4-20h, again pH is transferred to 6.0-8.0, stir 0.2-2h, centrifuge washing is dry, promptly gets.
2. a kind of environmental responsibility mesoporous silicon nanometer particle process method according to claim 1 is characterized in that: the template in the said step (1) is a cetyl trimethyl ammonium bromide.
3. a kind of environmental responsibility mesoporous silicon nanometer particle process method according to claim 1 is characterized in that: the particle diameter of the mesoporous silicon in the said step (1) is 20-1000nm, and specific surface area is 500-1500m
2/ g, average pore size is 2-50nm.
4. a kind of environmental responsibility mesoporous silicon nanometer particle process method according to claim 1 is characterized in that: the anionic polyelectrolyte in the said step (3) is maleic anhydride sodium salt, sodium alginate, kayexalate, polyacrylic acid, polymethylacrylic acid or polyacrylamide; Cationic polyelectrolyte is chitosan, polyallylamine hydrochlorate, PDDA or heparin.
5. a kind of environmental responsibility mesoporous silicon nanometer particle process method according to claim 1 is characterized in that: anionic polyelectrolyte or cationic polyelectrolyte solution concentration in the said step (3) are 0.2-2mg/mL.
6. a kind of environmental responsibility mesoporous silicon nanometer particle process method according to claim 1 is characterized in that: the bioactive molecule in the said step (4) is somatomedin, gene, enzyme or cancer therapy drug.
7. a kind of environmental responsibility mesoporous silicon nanometer particle process method according to claim 6 is characterized in that: said somatomedin is phosphoric acid sphingol, VEGF, basic fibroblast growth factor or epidermal growth factor.
8. a kind of environmental responsibility mesoporous silicon nanometer particle process method according to claim 6 is characterized in that: said gene is microRNA, disturb little RNA, messenger RNA, transfer RNA or ribosomal RNA.
9. a kind of environmental responsibility mesoporous silicon nanometer particle process method according to claim 6 is characterized in that: said enzyme is pectase, protease, lipase, catalase or amylase.
10. a kind of environmental responsibility mesoporous silicon nanometer particle process method according to claim 6 is characterized in that: said anticarcinogen is amycin, dexamethasone, lomustine, carmustine, dexrazoxane, fludarabine phosphate, capecitabine, paclitaxel, replaces lucky Austria, oxaliplatin, docetaxel, vinorelbine, elemene or hydroxy camptothecin.
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